Untargeted Metabolomics of Bladder Tissue using Liquid Chromatography and Quadrupole Time-of-Flight Mass Spectrometry for Cancer Biomarker Detection

Bladder cancer (BC) ranks among the most common cancers globally, with an increasing occurrence, particularly in developed nations. Utilizing tissue metabolomics presents a promising strategy for identifying potential biomarkers for cancer detection. In this study, we utilized ultra-high-performance liquid chromatography coupled with ultra-high-resolution mass spectrometry (UHPLC-UHRMS), incorporating both C18-silica and HILIC columns, to comprehensively analyze both polar and non-polar metabolite profiles in tissue samples from 99 patients with bladder cancer. By utilizing an untargeted approach with external validation, we identified twenty-five tissue metabolites that hold promise as potential indicators of BC. Furthermore, twenty-five characteristic tissue metabolites that exhibit discriminatory potential across bladder cancer tumor grades, as well as thirty-nine metabolites that display correlations with tumor stages were presented. Receiver-Operating Characteristics analysis demonstrated high predictive power for all types of metabolomics data, with area under the curve (AUC) values exceeding 0.966. Notably, this study represents the first report in which human bladder normal tissues adjacent to cancerous tissues were analyzed using UHPLC-UHRMS. These findings suggest that the metabolite markers identified in this investigation could serve as valuable tools for the detection and monitoring of bladder cancer stages and grades.